Department of Cell Biology and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Liangzhu Laboratory, Zhejiang University Medical Center, Hangzhou, China.
Nat Commun. 2022 Nov 28;13(1):7329. doi: 10.1038/s41467-022-35041-7.
Technologies for gene activation are valuable tools for the study of gene functions and have a wide range of potential applications in bioengineering and medicine. In contrast to existing methods based on recruiting transcriptional modulators via DNA-binding proteins, we developed a strategy termed Narta (nascent RNA-guided transcriptional activation) to achieve gene activation by recruiting artificial transcription factors (aTFs) to transcription sites through nascent RNAs of the target gene. Using Narta, we demonstrate robust activation of a broad range of exogenous and endogenous genes in various cell types, including zebrafish embryos, mouse and human cells. Importantly, the activation is reversible, tunable and specific. Moreover, Narta provides better activation potency of some expressed genes than CRISPRa and, when used in combination with CRISPRa, has an enhancing effect on gene activation. Quantitative imaging illustrated that nascent RNA-directed aTFs could induce the high-density assembly of coactivators at transcription sites, which may explain the larger transcriptional burst size induced by Narta. Overall, our work expands the gene activation toolbox for biomedical research.
基因激活技术是研究基因功能的有用工具，在生物工程和医学中有广泛的潜在应用。与现有的基于 DNA 结合蛋白募集转录调节剂的方法不同，我们开发了一种称为 Narta（新生 RNA 指导的转录激活）的策略，通过新生的靶基因 RNA 将人工转录因子（aTF）募集到转录位点，从而实现基因激活。使用 Narta，我们在各种细胞类型中，包括斑马鱼胚胎、小鼠和人类细胞中，展示了广泛的外源和内源基因的强大激活。重要的是，这种激活是可逆的、可调的和特异性的。此外，Narta 提供了一些表达基因的更好的激活效力比 CRISPRa，并且当与 CRISPRa 结合使用时，对基因激活具有增强作用。定量成像表明，新生 RNA 指导的 aTF 可以诱导转录位点处共激活因子的高密度组装，这可能解释了 Narta 诱导的更大转录爆发大小。总的来说，我们的工作为生物医学研究扩展了基因激活工具包。
